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High-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography

High-throughput X-ray crystal structures of protein–ligand complexes are critical to pharmaceutical drug development. However, cryocooling of crystals and X-ray radiation damage may distort the observed ligand binding. Serial femtosecond crystallography (SFX) using X-ray free-electron lasers (XFELs)...

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Autores principales: Moreno-Chicano, Tadeo, Ebrahim, Ali, Axford, Danny, Appleby, Martin V., Beale, John H., Chaplin, Amanda K., Duyvesteyn, Helen M. E., Ghiladi, Reza A., Owada, Shigeki, Sherrell, Darren A., Strange, Richard W., Sugimoto, Hiroshi, Tono, Kensuke, Worrall, Jonathan A. R., Owen, Robin L., Hough, Michael A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6830213/
https://www.ncbi.nlm.nih.gov/pubmed/31709063
http://dx.doi.org/10.1107/S2052252519011655
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author Moreno-Chicano, Tadeo
Ebrahim, Ali
Axford, Danny
Appleby, Martin V.
Beale, John H.
Chaplin, Amanda K.
Duyvesteyn, Helen M. E.
Ghiladi, Reza A.
Owada, Shigeki
Sherrell, Darren A.
Strange, Richard W.
Sugimoto, Hiroshi
Tono, Kensuke
Worrall, Jonathan A. R.
Owen, Robin L.
Hough, Michael A.
author_facet Moreno-Chicano, Tadeo
Ebrahim, Ali
Axford, Danny
Appleby, Martin V.
Beale, John H.
Chaplin, Amanda K.
Duyvesteyn, Helen M. E.
Ghiladi, Reza A.
Owada, Shigeki
Sherrell, Darren A.
Strange, Richard W.
Sugimoto, Hiroshi
Tono, Kensuke
Worrall, Jonathan A. R.
Owen, Robin L.
Hough, Michael A.
author_sort Moreno-Chicano, Tadeo
collection PubMed
description High-throughput X-ray crystal structures of protein–ligand complexes are critical to pharmaceutical drug development. However, cryocooling of crystals and X-ray radiation damage may distort the observed ligand binding. Serial femtosecond crystallography (SFX) using X-ray free-electron lasers (XFELs) can produce radiation-damage-free room-temperature structures. Ligand-binding studies using SFX have received only modest attention, partly owing to limited beamtime availability and the large quantity of sample that is required per structure determination. Here, a high-throughput approach to determine room-temperature damage-free structures with excellent sample and time efficiency is demonstrated, allowing complexes to be characterized rapidly and without prohibitive sample requirements. This yields high-quality difference density maps allowing unambiguous ligand placement. Crucially, it is demonstrated that ligands similar in size or smaller than those used in fragment-based drug design may be clearly identified in data sets obtained from <1000 diffraction images. This efficiency in both sample and XFEL beamtime opens the door to true high-throughput screening of protein–ligand complexes using SFX.
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spelling pubmed-68302132019-11-08 High-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography Moreno-Chicano, Tadeo Ebrahim, Ali Axford, Danny Appleby, Martin V. Beale, John H. Chaplin, Amanda K. Duyvesteyn, Helen M. E. Ghiladi, Reza A. Owada, Shigeki Sherrell, Darren A. Strange, Richard W. Sugimoto, Hiroshi Tono, Kensuke Worrall, Jonathan A. R. Owen, Robin L. Hough, Michael A. IUCrJ Research Papers High-throughput X-ray crystal structures of protein–ligand complexes are critical to pharmaceutical drug development. However, cryocooling of crystals and X-ray radiation damage may distort the observed ligand binding. Serial femtosecond crystallography (SFX) using X-ray free-electron lasers (XFELs) can produce radiation-damage-free room-temperature structures. Ligand-binding studies using SFX have received only modest attention, partly owing to limited beamtime availability and the large quantity of sample that is required per structure determination. Here, a high-throughput approach to determine room-temperature damage-free structures with excellent sample and time efficiency is demonstrated, allowing complexes to be characterized rapidly and without prohibitive sample requirements. This yields high-quality difference density maps allowing unambiguous ligand placement. Crucially, it is demonstrated that ligands similar in size or smaller than those used in fragment-based drug design may be clearly identified in data sets obtained from <1000 diffraction images. This efficiency in both sample and XFEL beamtime opens the door to true high-throughput screening of protein–ligand complexes using SFX. International Union of Crystallography 2019-10-10 /pmc/articles/PMC6830213/ /pubmed/31709063 http://dx.doi.org/10.1107/S2052252519011655 Text en © Tadeo Moreno-Chicano et al. 2019 http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.http://creativecommons.org/licenses/by/4.0/
spellingShingle Research Papers
Moreno-Chicano, Tadeo
Ebrahim, Ali
Axford, Danny
Appleby, Martin V.
Beale, John H.
Chaplin, Amanda K.
Duyvesteyn, Helen M. E.
Ghiladi, Reza A.
Owada, Shigeki
Sherrell, Darren A.
Strange, Richard W.
Sugimoto, Hiroshi
Tono, Kensuke
Worrall, Jonathan A. R.
Owen, Robin L.
Hough, Michael A.
High-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography
title High-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography
title_full High-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography
title_fullStr High-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography
title_full_unstemmed High-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography
title_short High-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography
title_sort high-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6830213/
https://www.ncbi.nlm.nih.gov/pubmed/31709063
http://dx.doi.org/10.1107/S2052252519011655
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